Investigation of the Nucleotide Sequence Variations of Genes mazE and mazF in Escherichia coli Bacteria Isolated from Clinical Sources
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Abstract
Gene pairs known as toxin‒antitoxin (TA) modules are crucial for the control of bacteria in response to environmental stressors. The MazF toxin and the associated MazE antitoxin are encoded by the mazEF module. The present study aimed, for the first time, to determine the relationship between multidrug resistance in E. coli isolates and nucleotide sequence variation in one of the toxin-antitoxin family genes in uropathogenic Iraqi isolates. Among the thirty isolates, only 11 (36.6%) were identified as E. coli on the basis of microbiological tests and biochemical tests and then confirmed by molecular identification via the dinB gene. Antimicrobial resistance tests were performed via the Kirby–Bauer method, and amplification of the genes mazE and mazF via PCR was performed on 7 (63.6%) isolates. The results revealed the incidence of some substitution gene mutations from 100–98.61% identity with the reference sequence gene of the global strain with accession no. (mazEF-947252) by alignment in NCBI by Sanger sequencing, and the phylogenetic tree was drawn via Geneious software. Analysis of the phylogenetic tree revealed five different groups of variation. The code isolates (Escherichia coli ZSSh33 mazE gene, ZSSh40 mazE gene and ZSSh9 mazF gene) were chosen and documented in NCBI as new Iraqi isolates and accepted under these accession numbers (LC600699, LC600700 and LC600698, respectively).
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References
1. Tripathi A, Dewan PC, Siddique SA and Varadarajan R. MazF-induced growth inhibition and persister generation in Escherichia coli. J. biol. chem.2014;289(7):4191–4205. https://doi.org/10.1074/jbc.M113.510511
2. Choi W, Yamaguchi Y, Park JY, Park SH, Lee HW, Lim BK, Otto M, Inouye M, Yoon MH and Park JH.Functional Characterization of the mazEF Toxin-Antitoxin System in the Pathogenic Bacterium Agrobacterium tumefaciens. Microorg.2021: 9(5):1107. https://doi.org/10.3390/microorganisms9051107
3. Kamruzzaman M, Wu AY and Iredell JR. Biological Functions of Type II Toxin-Antitoxin Systems in Bacteria. Microorg..2021: 9(6):1276. https://doi.org/10.3390/microorganisms9061276
4. Al-Shimmary SM. Molecular identification and prevalence of some virulence genes among Pseudomonas aeruginosa isolated from Iraqi patients. Int. J. Pharma.Res.2020.Supplementary Issue 1: 1542.
5. Harms A, Brodersen DE, Mitarai N and Gerdes K. Toxins, Targets, and Triggers: An Overview of Toxin-Antitoxin Biology. Mole. cell.
2018: 70(5):768–784. https://doi.org/10.1016/j.molcel.2018.01.003
6. Lee KY, and Lee BJ. Structure, Biology, and Therapeutic Application of Toxin-Antitoxin Systems in Pathogenic Bacteria. Tox. 2016: 8(10):305. https://doi.org/10.3390/toxins8100305
7. Yang QE and Walsh TR. Toxin-antitoxin systems and their role in disseminating and maintaining antimicrobial resistance. FEMS microbial. rev.2017 41(3):343–353. https://doi.org/10.1093/femsre/fux006
8. Qiu J, Zhai Y, Wei M, Zheng C and Jiao X. Toxin–antitoxin systems: Classification, biological roles, and, application. Microbiol. Res.2022:264:127159. https://doi.org/10.1016/j.micres.2022.127159
9. Anderson C, Johnson TR, Case CL, Cappuccino JG and Sherman N.Great Adventures in the Microbiology Labratory 8th (ed). Pearson Custom Publishing:2015:175–176.
10. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing; 31st (ed.),2021 CLSI supplement M100 (ISBN 978-1-68440-104-8 (Print), and ISNB 978-168440-105-5 (Electronic) were used. Clinical and Laboratory Standard Institute, USA, https://www.treata.academy/wp-content/uploads/2021/03/CLSI-31-2021.pdf
11. Shehab ZH, Sweedan EG and Flayyih MT. Evaluation of the effects of Allium sativum (garlic) oil on the expression of the mazE and mazF genes in E. coli clinical isolates. Biochem. Cell. Arch .2021:21(1):721-726.
12. Mohaisen SH, Ali MH, Shehab ZH, Al-Mayyahi A and Abdulhassan AJA. Effect of Ultraviolet light on the expression of icaD gene in Staphylococcus aureus local isolates in Iraq. Arch. Razi Instit.2021;76(5):1221-1227.
13. Henrikus SS, Wood EA, McDonald JP, Cox MM, Woodgate R, Goodman MF, van Oijen AM, and Robinson A.DNA polymerase IV primarily operates outside of DNA replication forks in Escherichia coli. PLOS Gen.2018: 14(1): e1007161.https://doi.org/10.1371/journal.pgen.1007161
14. Al-Shimmary SM , Almohaidi AMS, Al-Qaysi SAS, Mohamed NS. Improvement Rapid Molecular Detection of Pseudomonas aeruginosa infected some Iraqi Patients and It is antimicrobial susceptibility. Res. J. Pharm. Biol. Chem. Sci.., 2016; 7(5)1256-1264.
15. Nasif SH, Ali, MR, and Alsakini, AH.Characterization of Uropathogenic Escherichia coli by Phylogenetic Grouping, Integron, and Antibiotic Resistance Properties. Trop J Nat Prod Res. 2024; 8(4): 70357041. https://doi.org/10.26538/tjnpr/v8i4.41
16. Redha MA, Al Sweih N and Albert MJ. Multidrug-Resistant and Extensively Drug-Resistant Escherichia coli in Sewage in Kuwait: Their Implications. Microorg. 2023; 11(10):2610. https://doi.org/10.3390/microorganisms11102610
17. AL-Lami, RAH, Al-Hayanni HAS, Shehab ZH. Molecular Investigation of Some Beta-lactamase Genes by PCR andDNA Sequencing Techniques in clinical Escherichia coli. Iraqi J. Sci. 2022;63(10: 4205–4212. doi: 10.24996/ijs.2022.63.10.7
18. Adenike O and Ayansina AD. Occurrence of Extended Spectrum Beta Lactamase (ESBL) Producing Gram-Negative Bacteria in Wastewaters from Selected Hospitals in Ibadan, Oyo State, Nigeria. Trop J Nat Prod Res.2022; 6(5): 826-830. https://www.tjnpr.org/index.php/home/article/view/69
19. Rahminiwati M, Batubara, I, Nurcholis, W, Safinatunajah RR, Sari FE and Sulistiyono FD. Antibacterial and Antibiofilm Properties of Jicama (Pachyrhizus erosus) Seed Extract against Streptococcus mutans: Identification of Key Bioactive Compounds Using Bioautography and LCMS/MS Analysis. Trop J Nattat Prod Res.2025;9(1), 97-103. https://doi.org/10.26538/tjnpr/v9i1.14
20. Jurėnas D, Fraikin, N, Goormaghtigh F and Van Melderen L. Biology and Evolution of Bacterial Toxin–Antitoxin Systems. Nat. Rev. Microbiol.2022; 20:335–350.
21. Norton JP and Mulvey MA. Toxin-antitoxin systems are important for niche-specific colonization and stress resistance of uropathogenic Escherichia coli. PLoS Pathog. 2012; 8:e1002954. doi: 10.1371/journal.ppat.1002954.
22. Bustamante, P.; Vidal, R. Repertoire and Diversity of Toxin-Antitoxin Systems of Crohn’s Disease-Associated Adherent-Invasive Escherichia coli. New Insight of T His Emergent E. coli Pathotype. Front. Microbiol. 2020;11:807. https://doi.org/10.3389/fmicb.2020.00807
23. Griffiths AJF, Miller JH, Suzuki DT, Lewontin RC and Gelbart WM .An Introduction to Genetic Analysis, 7th (ed.).2000,New York: WH Freeman.
24. Watford S and Warrington SJ. Bacterial DNA Mutations.2023: In StatPearls. StatPearls Publishing.